棉花黄萎病菌致萎毒素的分离纯化方法  

Analysis on physicochemical properties of toxins produced by three verticillium dahliae strains and it's purifying method

作　　者：陈旭升[1] 王祝鸣[2] 

机构地区：[1]江苏省农科院经济作物研究所,南京210014 [2]南京医科大学分子生物学研究所,南京210029

出　　处：《上海交通大学学报（农业科学版）》2002年第2期141-144,共4页Journal of Shanghai Jiaotong University(Agricultural Science)

摘　　要：利用快速蛋白质液相色谱技术 (Fast Protein Liquid Chromatography,FPLC) ,对棉花 3个黄萎病菌 (Verticilliumdahliae)菌系外泌毒素的某些理化性质进行了分析。结果显示 :黄萎病菌毒素有两个致萎峰 ,大分子峰 毒蛋白致萎力较弱 ,致萎症状属黄斑型 ;而小分子峰 毒蛋白其致萎力极强 ,致萎症状属青枯型。透析试验表明 ,可透 80 0 0～ 1 0 0 0 0 Da的透析袋会使小分子峰 丢失。热稳定试验表明 :峰 、峰 毒蛋白皆可耐 1 0 0℃高温水浴热煮而不发生沉淀 ,并保持致萎活性。根据以上研究结果 ,本文提出了黄萎病毒素分离纯化新方法。The toxic proteins secreted by three typical Verticillium dahliae strains T9?VD8 and SS-4 were separated by the technology of fast protein liquid chromatography (FPLC) into two wilting peaks. The first peak had a weak wilting ability showing yellowish wilt on cotton seedlings with normal root system and the second one resulted in strong wilting to cause severe greenish wilt. Furthermore, the second peaks' were lost after dialysed by the dialyser of MWCO 8 000-10 000 Da, So the dialyser or superfiltrator for molecular weight of 10 000 or more dalton was not suitable for desalting of wilting toxin secreted by V. dahliae. The collected peaks were tolerant of high temperatures. After kept in water bath at 100℃ for an hour, the crude extracting toxins from the three strains still had two wilting peaks through the gel column of Superose 12. According to the research result of toxin's physicochemical properties,the method of purifying crude extracting toxin from Verticillium dahliae was proposed .

关 键 词：棉花 黄萎病 黄萎病菌 毒素 FPLC 分离 纯化 

分 类 号：S435.621.2[农业科学—农业昆虫与害虫防治]